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Optimization of protectant, salinity and freezing condition for freeze-drying preservation of Edwardsiella tarda

文献类型： 外文期刊

作者： Yu, Yongxiang 1 ; Zhang, Zheng 2 ; Wang, Yingeng 2 ; Liao, Meijie 2 ; Li, Bin 2 ; Xue, Liangyi 1 ;

作者机构： 1.Ningbo Univ, Coll Marine Sci, Ningbo 315211, Zhejiang, Peoples R China

2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266071, Peoples R China

关键词： freeze-drying;mariculture;pathogen;Edwardsiella tarda;protectant;orthogonal test;interaction effect

期刊名称：JOURNAL OF OCEAN UNIVERSITY OF CHINA （ 影响因子：0.913； 五年影响因子：1.012 ）

ISSN： 1672-5182

年卷期： 2017 年 16 卷 5 期

页码：

收录情况： SCI

摘要： Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose (8.0%), skim milk (12.0%), sodium citrate (2.0%), serum (12.0%) and PVP (2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of NaCl was 10.0, 30.0 and between 5.0 and 10.0 g L-1 for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at -80A degrees C or -40A degrees C for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate (79.63%-82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy (SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.

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